Common Buffers, Media, and Stock Solutions

AbstractThis appendix describes the preparation of selected bacterial media and of buffers and reagents used in the manipulation of nucleic acids and proteins. Recipes for cell culture media and reagents are located elsewhere in the manual. Recipes Acids, concentrated stock solutions Ammonium acetate, 10 M Ammonium hydroxide, concentrated stock solution ATP, 100 mM BCIP, 5% (w/v) BSA (bovine serum albumin), 10% (100 mg/ml) Denhardt solution, 100× dNTPs: dATP, dTTP, dCTP, and dGTP DTT, 1 M EDTA, 0.5 M (pH 8.0) Ethidium bromide solution Formamide loading buffer, 2× Gel loading buffer, 6× HBSS (Hanks balanced salt solution) HCl, 1 M HEPES‐buffered saline, 2× KCl, 1 M LB medium LB plates Loading buffer 2‐ME, (2‐mercaptoethanol)50 mM MgCl2, 1 M MgSO4, 1 M NaCl, 5 M NaOH, 10 M NBT (nitroblue tetrazolium chloride), 5% (w/v) PCR amplification buffer, 10× Phosphate‐buffered saline (PBS), pH ∼7.3 Potassium acetate buffer, 0.1 M Potassium phosphate buffer, 0.1 M RNase a stock solution (DNase‐free), 2 mg/ml SDS, 20% SOC medium Sodium acetate, 3 M Sodium acetate buffer, 0.1 M Sodium phosphate buffer, 0.1 M SSC (sodium chloride/sodium citrate), 20× SSPE (sodium chloride/sodium phosphate/EDTA), 2